Api5500 triple quadrupole mass

Mass spectrometry detection was performed using an API5500 triple quadrupole mass (AB SCIEX, Framingham, MA, USA) equipped with an electrospray ionization (ESI) source operating in both positive and negative ion modes, and all data were acquired and analyzed by the Analyst 1.6.2 software. The ESI–MS spectra were acquired in the MRM. The parameters in the source were set as follows: GS1 flow: 55 L/min, GS2 flow: 55 L/min, and CUR flow: 40 L/min; gas temperature: 550 °C; pressure of the nebulizer of MS: 4500 V for the positive ion mode and −4500 V for the negative ion mode.

The mass spectrometry detection was performed using an API5500 triple quadrupole mass (AB SCIEX, Framingham, MA, USA). The MS was equipped with an electrospray ionization (ESI) source operating in MRM and under both positive and negative ion modes. The MS parameters were set as follows: gas temperature 550 °C; pressures of nebulizer of MS, 5500 V (positive) and −4500 V (negative); GSI flow 65 L/min; CUR flow 30 L/min and all MS data were acquired and analyzed using the Analyst 1.5.2 software (AB SCIEX, Framingham, MA, USA). The cone voltage and collision energy parameter of each compound were individually optimized.
The chromatographic analysis was performed on a Shimadzu SIL-20A XR system (Shimadzu, Kyoto, Japan), consisting of a binary solvent delivery system and an automatic sampler. A Synergi^TM Hydro-RP 100Å column (100 mm × 2.0 mm, 2.5 μm, (Phenomenex, Los Angeles, CA, USA) was used for eluting samples. The mobile phase was composed of water (A) and acetonitrile (B) using a gradient elution of 30%–52% B at 0–4 min, 52%–75% B at 4–8 min, 75%–90% B at 8–11 min, 90%–30% B at 11–15 min, 30% B at 15–17.10 min. The column temperature was 40 °C, the flow rate kept at 0.4 mL/min, and the sample injection volume was 1 μL.

Chromatographic analysis was performed on a Shimadzu SIL-20A XR system (Shimadzu, Kyoto, Japan), consisting of a binary solvent delivery system and an autosampler. Separation was performed on a Waters XBridge Amide (2.1 mm × 100 mm, 3.5 μm) column. The mobile phase was composed of water with 0.2% formic acid (A) and acetonitrile with 0.2% formic acid (B) using a gradient elution of 15% A at 0–2.5 min, 15%–50% A at 2.5–5 min, 50% A at 5–7 min, 50%–15% A at 7–8 min, 15% A at 8–11 min. The flow rate was 0.6 mL/min, and the column temperature was set at 30 °C.
Mass spectrometry detection was performed using an API5500 triple quadrupole mass (AB SCIEX, Framingham, MA, USA) equipped with an electrospray ionization (ESI) source operating in the negative ion mode. The ESI-MS spectra were acquired in the multiple reaction monitoring (MRM). The parameters in the source were set as follows: GS1 flow 55 L/min, GS2 flow 55 L/min, CUR flow 40 L/min; gas temperature 550 °C; pressure of nebulizer of MS −4500 V. All MS data were acquired using the Analyst 1.6.2 software to ensure mass accuracy and reproducibility.